Characterization of genes related to the efflux pump and porin in multidrug-resistant Escherichia coli strains isolated from patients with COVID-19 after secondary infection.

BACKGROUND: Escherichia coli (E. coli) is a multidrug resistant opportunistic pathogen that can cause secondary bacterial infections in patients with COVID-19. This study aimed to determine the antimicrobial resistance profile of E. coli as a secondary bacterial infection in patients with COVID-19 and to assess the prevalence and characterization of genes related to efflux pumps and porin. METHODS: A total of 50 nonduplicate E. coli isolates were collected as secondary bacterial infections in COVID-19 patients. The isolates were cultured from sputum samples. Confirmation and antibiotic susceptibility testing were conducted by Vitek 2. PCR was used to assess the prevalence of the efflux pump and porin-related genes in the isolates. The phenotypic and genotypic evolution of antibiotic resistance genes related to the efflux pump was evaluated. RESULTS: The E. coli isolates demonstrated high resistance to ampicillin (100%), cefixime (62%), cefepime (62%), amoxicillin-clavulanic acid (60%), cefuroxime (60%), and ceftriaxone (58%). The susceptibility of E. coli to ertapenem was greatest (92%), followed by imipenem (88%), meropenem (86%), tigecycline (80%), and levofloxacin (76%). Regarding efflux pump gene combinations, there was a significant association between the acrA gene and increased resistance to levofloxacin, between the acrB gene and decreased resistance to meropenem and increased resistance to levofloxacin, and between the ompF and ompC genes and increased resistance to gentamicin. CONCLUSIONS: The antibiotics ertapenem, imipenem, meropenem, tigecycline, and levofloxacin were effective against E. coli in patients with COVID-19. Genes encoding efflux pumps and porins, such as acrA, acrB, and outer membrane porins, were highly distributed among all the isolates. Efflux pump inhibitors could be alternative antibiotics for restoring tetracycline activity in E. coli isolates.

Accessory Gene Regulator (agr) group polymorphisms in methicillin-resistant Staphylococcus aureus and its association with biofilm formation.

 Methicillin-resistant Staphylococcus aureus (MRSA) is one of the main causes of community- and hospital-acquired infections. The expression of virulence genes in S. aureus is arranged by regulators like the accessory gene regulator (agr). The present study aims to estimate phenotypic characteristics of S. aureus and investigate the occurrence of agr genes with their correlation to biofilm formation. In this study, 34 MRSA strains out of 100 S. aureus isolates were recovered in a variety of clinical samples. Phenotypic characterization and ability of biofilm formation were assessed. About 8(24%) of isolates were biofilm producers. The percentages of biofilm production among isolates were 3(37.5%), 2(25%), 3(37.5%) as strong, moderate, and weak, respectively. Furthermore, the resistance rates for all antibiotics were higher in biofilm producers and 76% of the isolates were staphyloxanthin producers, around 82% of the strains showed resistance to H2O2. Hemolytic activity was detected in 74% of the total isolates. The activity of the protease enzyme was 68%. The lipase enzyme was active in 79% of the tested S. aureus isolates. The majority of isolates were established to be agrI 84%, followed by agrII 53%, agrIII 32%, and 30% of the isolates have agr IV. Our study indicated that the majority of MRSA isolates were non-biofilm producers and the agr I is the most dominant type. Thus, agr I is not correlated with biofilm production.

Antibacterial and Antioxidant Study of New Pharmaceutical Formulation of Didecyldimethylammonium Bromide Via Pharmaceutical Deep Eutectic Solvents (PDESs) Principle.

Combination therapies have been studied by many researchers using different techniques and methods to solve some solid drug problems and improve more effective treatments for humans and animals. One of the more significant findings to emerge from this study is that the combination of pharmaceutical agents by using pharmaceutical deep eutectic solvents (PDESs) in order to produce dual action drugs and reduce the drug resistance. The major objective of this study was to investigate the dual functionality of drugs (antioxidant and antibacterial activity) via the principle of PDESs. The produced PDESs were characterized via different techniques, namely differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), and UV-Vis spectrophotometry. We herein tested a panel of novel liquid formulations of didecyldimethylammonium bromide (DDMAB) against a selection of pathogenic bacteria, classifying their spectrum of activity against Gram-positive and Gram-negative bacteria. The current study found that the PDESs can be used to produce drugs with dual functionalities. The produced PDES from (ascorbic acid: DDMAB) exhibits stronger antibacterial activity against Gram-positive Staphyloccocus aureus and Staphyloccocus epidermidis than gram negatives. One of the most interesting PDESs studied in this research was that of DDMAB and ascorbic acid. This forms a eutectic which is far from the solid drugs issues and shows dual functionality like antibacterial and antioxidant activity. This study has found that there is a correlation between the molecular docking study and the biological activities of the combined drugs.

Down-regulation of abaI, abaR, Bap and OmpA genes in Acinetobacter baumannii by ethanol extract of Glycyrrhiza glabra after toxicity assessment.

Acinetobacter baumannii is a pathogen that has caused rising concerns within healthcare facilities in recent years. As antibiotic overuse and resistance rise, natural remedies with the potential have received attention as antibiotics that might have fewer side effects and lower resistance. Glycyrrhiza glabra was used to investigate its effects on A. baumannii's quorum sensing and biofilm production abilities. In this study, the toxicity assessment of Glycyrrhiza glabra L. extract on rats, the phytochemical analysis and the quantitative measurement for the association of the biofilm reduction with the active components in the plant was determined. The results indicated ciprofloxacin and gentamicin were the most effective antibiotics and that various capabilities of biofilm-productions were demonstrated, only four percent of the samples established robust biofilm, while 40% to 56% demonstrated weak to moderate biofilm production, respectively. Phytochemical qualitative testing of ethanol leaf extracts from Glycyrrhiza glabra showed the existence of flavonoids, alkaloids, phenolic, tannic acid, and terpenoids, but no saponins. Assessment of toxicity revealed a low hazard, with an LD50 of 4.95 g/Kg. Our results showed that the extract's SICs elucidated a substantial quantitative decrease in biofilm production by the bacterial isolates, including the reference ATCC strain, which is known to be a potent biofilm producer. As a conclusion, biofilm creation in Acinetobacter baumannii has been shown to be greatly reduced by G. glabra extract.

Diversity of biofilm-specific antimicrobial resistance genes in Pseudomonas aeruginosa recovered from various clinical isolates.

Background and Objectives: The resistance of Pseudomonas aeruginosa to antibiotics offers a significant challenge in the treatment of patients. This study aimed to investigate the antimicrobial resistance profile, biofilm-specific antimicrobial resistance genes, and genetic diversity of P. aeruginosa recovered from clinical samples. Materials and Methods: Totally 47 non-duplicate isolates of P. aeruginosa were recovered from various clinical samples. toxA, algD, ndvB, and tssC1 genes were detected in biofilm-producing isolates. The DNA sequences of the toxA and tssC1 genes were analyzed, by creating phylogenetic trees. Results: The findings revealed that 30 (63.8%) of the isolates tested positive for Extended spectrum ß-lactamase (ESBL), whereas 31 (65.9%) tested positive for Metallo-ß-lactamase (MBL) and all of the isolates presented the toxA genes, and 19.1%,17%, 6.3% presented by algD, ndvB and tssC1 genes. Besides, the phylogenetic trees of the toxA and tssC1 gene isolates suggested a genotype that was closely aligned with others. Gene sequencing similarity revealed 99% identity with other isolates deposited in GenBank. Conclusion: The occurrence of toxA was most prevalent. One isolate was recorded as a novel isolate in the global gene bank as a locally isolated strain from the city of Erbil that has never been identified in global isolates due to genetic variation.

DLX6-AS1: A Long Non-coding RNA With Oncogenic Features.

Long non-coding RNAs (lncRNAs) are a heterogeneous group of ncRNAs with characteristic size of more than 200 nucleotides. An increasing number of lncRNAs have been found to be dysregulated in many human diseases particularly cancer. However, their role in carcinogenesis is not precisely understood. DLX6-AS1 is an lncRNAs which has been unveiled to be up-regulated in various number of cancers. In different cell studies, DLX6-AS1 has shown oncogenic role via promoting oncogenic phenotype of cancer cell lines. Increase in tumor cell proliferation, migration, invasion, and EMT while suppressing apoptosis in cancer cells are the effects of DLX6-AS1 in development and progression of cancer. In the majority of cell experiment, mediator miRNAs have been identified which are sponged and negatively regulated by DLX6-AS1, and they in turn regulate expression of a number of transcription factors, eventually affecting signaling pathways involved in carcinogenesis. These pathways form axes through which DLX6-AS1 promotes carcinogenicity of cancer cells. Xenograft animal studies, also have confirmed enhancing effect of DLX6-AS1 on tumor growth and metastasis. Clinical evaluations in cancerous patients have also shown increased expression of DLX6-AS1 in tumor tissues compared to healthy tissues. High DLX6-AS1 expression has shown positive association with advanced clinicopathological features in cancerous patients. Survival analyses have demonstrated correlation between high DLX6-AS1 expression and shorter survival. In cox regression analysis, DLX6-AS1 has been found as an independent prognostic factor for patients with various types of cancer.